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Establishment of Indirect Competitive ELISA for the Detection of Zearalenone Produced by Fusarium sp. (Fusarium속이 생성하는 zearalenone 측정을 위한 Indirect Competitive ELISA의 확립)

  • 강성조;정덕화
    • Journal of Food Hygiene and Safety
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    • v.13 no.4
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    • pp.419-424
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    • 1998
  • An enzyme-linked immunosorbent assay (ELISA) was established for the detection of zearalenone by using monoclonal antibodies produced by Z-M-26 hybridoma cells when injected into a mouse and zearalenone-oxime-OV A conjugate. Zearalenone-oxime-OV A conjugates were diluted with carbonyl buffer, coated to 96 well microtiter plates at $4^{\circ}C$ overnight and blocked with 1% BSA overnight. One thousand times diluted antibody solution together with standard zearalenone or sample was added to 96-well microtiter plates and stood overnight. A secondary antibody conjugated with HRP was added and an hour later, enzyme substrate (TMBZ) solution was added for color develpment. Mter 30 minutes, coloring reaction was terminated by adding 2 N $H_2S0_4$ and the O.D. was measured at 450 nm. Detection range of this method was about 0.1~100 ppb. The established indirect competitive ELISA method was suitable for a rapid and effective analysis of zearalenone in agricultural products.

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The Electrochemical Characteristics of 9-methyl-2,3,6,7-tetramethoxyfluorene in CH3CN and CH2Cl2/TFA/TFAn-solvent Mixture (CH3CN과 CH2Cl2/TFA/TFAn-혼합용매계에서의 9-methyl-2,3,6,7-tetramethoxyfluorene의 전기화학적 특성)

  • Kim, Duk-Hyun;Kim, Su-Jin
    • Applied Chemistry for Engineering
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    • v.10 no.2
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    • pp.196-200
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    • 1999
  • The electrochemical properties of 9-methyl-2,3,6,7-tetramethoxyfluorene have been investigated by cyclic voltammetry in acetonitrile, dichloromethane, trifluoroacetic acid (TFA) and trifluoroacetic acid anhydride (TFAn). The first charge transfer for the compound in $CH_3CN$ appeared to be a quasi-reversible one-electron step. The second oxidation step from cation to dication was irreversible. However, the oxdition of the compound in a mixture of solvents containing $CH_2Cl_2$, TFA and TFAn was reversible for both the first and second charge transfer reactions. Since the electrolytic products display a darkblue color and can be stabilized in the solvent mixture, they may be used as an electrochromic material.

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Development of Enzyme Linked Immunosorbent Assay for Determination of Zearalenone in Animal Feeds (가축사료 중 Zearalenone 분석을 위한 Enzyme Linked Immunosorbent Assay법의 개발)

  • 하정기;정덕화;김성영
    • Journal of Food Hygiene and Safety
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    • v.6 no.3
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    • pp.111-117
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    • 1991
  • We examined to develop the enzyme linked immunosorbent assay (ELISA) for determination of zearalenone in animal feeds. Zearalenone was first converted to 6'-(carboxymethyl) zearalenone oxime(zearalenone oxime) to get a coupling site and then conjugated to bovine serum albumin(BSA) for use as immunogen and to horseradish peroxidase(HRP) for use as enzyme marker. Antibody against zearalenone was obtained after 11 weeks of immunization of rabbit with zearalenone oxime-BSA. Cross reactivity of the antibody with ${\alpha}-zearalanol,\;{\beta}-zearalenol,\;{\alpha}-zearalanol\;and\;{\beta}-zearalanol$ were 168, 46, 26 and 20% respectiviely. A simple procedure was devised for the screening of zearalenone in feeds using ELISA. Feeds samples(5g) were extracted by blending with 25 ml of methanol-phospate butTered saline-dimethylformate(70 : 29 : 1) and the extract was filtered and aqueous filterate analyzed. It took only 1 hours to do whole procedure for the analysis of zearalenone in feeds by the direct competitive ELISA, and detectable limit was 1-100 ppb. Using this procedure, only 4 of 24 feed samples showed positive results with 3.93-7.43 ppb levels.

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Electrochemical Characteristic on Hydrogen Intercalation into the Interface between Electrolyte of the 0.1N H2SO4and Amorphous Tungsten Oxides Thin Film Fabricated by Sol-Gel Method (졸-겔법으로 제조된 비정질의 텅스텐 산화물 박막과 황산 전해질 계면에서 일어나는 수소의 층간 반응에 대한 전기화학적 특성)

  • Kang, Tae-Hyuk;Min, Byoung-Chul;Ju, Jeh-Beck;Sohn, Tae-Won;Cho, Won-Il
    • Applied Chemistry for Engineering
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    • v.7 no.6
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    • pp.1078-1086
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    • 1996
  • The peroxo-polytungstic acid was formed by the direct reaction of tungsten powder with the hydrogen peroxide solution. Peroxo-polytungstic powder were prepared by rotary evaporator using the fabricated on to ITO coated glass as substrate by dip-coating method using $2g/10mL(W-IPA/H_2O)$ sol solution. A substrate was dipped into the sol solution and after a meniscus had settled, the substrate was withdrawn at a constant rate of the 3mm/sec. Thicker layer could be built up by repeated dipping/post-treatment 15 times cycles. The layers dried at the temperature of $65{\sim}70^{\circ}C$ during the withdrawn process, and then tungsten oxides thin film was formed by final heating treatment at the temperature of $230{\sim}240^{\circ}C$ for 30min. A linear rotation between the thickness of thin film and the number of dipping/post-treatment cycles for tungsten oxides thin films made by dip-coating was found. The thickness of thin film had $60{\AA}$ after one dipping. From the patterns of XRD, the structure of tungsten oxides thin film identified as amorphous one and from the photographs of SEM, the defects and the moderate cracks were observed on the tungsten oxides thin film, but the homogeneous surface of thin films were mostly appeared. The electrochemical characteristic of the $ITO/WO_3$ thin film electrode were confirmed by the cyclic voltammetry and the cathodic Tafel polaization method. The coloring bleaching processes were clearly repeated up to several hundreds cycles by multiple cyclic voltammetry, but the dissolved phenomenon of thin film revealed in $H_2SO_4$ solution was observed due to the decrease of the current densities. The diffusion coefficient was calculated from irreversible Randles-Sevick equation from the data obtained by the cyclic voltammetry with various scan rates.

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A Recombinant Microbial Biosensor for Cadmium and Lead Detection (카드뮴 및 납 검출을 위한 재조합 미생물 바이오센서)

  • Shin, Hae Ja
    • Journal of Life Science
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    • v.26 no.5
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    • pp.503-508
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    • 2016
  • Biosensors have been used as first-step monitoring tools to detect on-site samples in a simple and cost-effective manner. Numerous recombinant microbial biosensors have been exploited for monitoring on-site toxic chemicals and biological signals. Herein, a recombinant microbial biosensor was constructed for monitoring cadmium. The cadmium responding cadC regulatory gene and it’s promoter from Staphylococcus aureus was amplified through PCR, fused with the lacZ gene, and transformed into Escherichia coli BL21 (DE3) cells. In the presence of cadmium, the biosensor cells express β-galactosidase showing red color development with chlorophenol red β-galactopyranoside (CPRG) as the enzymatic substrate. The biosensor cells showed the best β-galactosidase activity after 3 hr induction with cadmium at pH 5 and a detection range from 0.01 μM to 10 mM cadmium with a linearity from 0.01 to 0.1 μM cadmium (y = 0.98 x + 0.142, R2 = 0.98). Among the heavy metals, cadmium and lead showed good responses, tin and cobalt showed medium responses, and mercury and copper showed no responses. The biosensor cells showed good responses to several waste waters similar to buffer solution, all spiked with cadmium. The biosensor described herein could be applied for on-site cadmium monitoring in a simple and cost-effective manner without sample pretreatments.

1,3-bisdicyanovinylindane 색소를 이용한 선택적 $Hg^{2+}$ 감지 특성

  • Kim, Su-Ho;Kim, Young-Sung;Kim, Sung-Hoon;Son, Young-A
    • Proceedings of the Korean Society of Dyers and Finishers Conference
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    • 2009.11a
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    • pp.19-20
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    • 2009
  • 최근 화학, 물리, 생명과학, 전기, 전자등의 다양한 분야에 활발하게 연구가 이루어지고 있는 초분자 화학은 선택적 분자인지를 위한 효율적인 골격구조와 나아가 다양한 계에 응용할 수 있다. 초분자 화학의 분자인지 과정의 특징은 일반적으로 수용체 (receptor 혹은 host)가 목표가 되는 기질 (substrate, analyte, 혹은 guest)에 대하여 선택적으로 식별하고 반응하는 것이다. 비공유 결합성 상호작용에 의하여 이루어지는 초분자 화학의 분자 인지 과정의 특징은 일반적으로 수용체 (receptor 혹은 host)가 목표가 되는 기질 (substrate, analyte, 혹은 guest)에 대하여 선택적으로 식별하고 반응하는 것이다. 이는 공유결합을 이용하는 분자화학과는 차별화 된 것이다. 수용체는 간단한 구조의 화합물 및 금속 이온들과 같은 기질과 가역적으로 상호 작용할 수 있는 착물을 형성한다. 최근들어 급격한 산업화가 진행되어 환경문제가 심각하게 대두 되어져 왔고, 그 중에서 특히 수은이나 카드뮴에 의한 질병, 납에 의한 중독 등 중금속에 의한 오염이 크게 나타남에도 불구하고, 현재 그러한 중금속을 검출함에 있어 많은 비용과 시간이 드는 문제점이 있다. 또한 우리에게 이로운 금속은 효율적 분석을 통해 환경계와 의료계에 많은 도움을 줄 것으로 사례되므로 화학센서 기술의 개발은 절실히 요구되어지고 있다. 이에 새로운 1,3-bisdicyanovinylindane 을 통해 $Hg^{2+}$ 금속의 감지 여부 알아보고, 그 특성을 파악하고자 한다. 1,3-indandion (2.18g, 14.9mmol), malononitrile (2.95g, 44.7mmol), ethanol 50ml를 20분간 상온에서 용해시킨다. 후에 sodium trihydrate acetate(3.05g)을 첨가한 후 5시간 동안 환류반응 시킨다. 이 과정에서 얻어진 용액을 필터과정을 통하여 에서 합성 반응 중에 생성된 불순물(1,3-dicyanovinylindane-1-one, monocondensation)을 제거한다. 필터과정을 통해 걸러진 미 반응 물질을 제�G 용반응욕액을 증류수(100ml)를 이용하여 희석시키고 난 후 염산을 이용, 산성화 시켜 고체 생성물을 얻어낸다. 이렇게 생성된 고체 생성물은 다시 필터 및 건조를 통하여 회색의 고체 화합물을 얻어낸다. 1,3-bisdicyanovinylindane과 금속이온에 대한 감응도를 확인하기 위하여 metanol/water(1:2)을 용매로 하여 금속이온의 농도를 변화시켜 발색특성을 살펴보았다. 본 색소화합물과 Hg2+에 대한 UV 흡광도 변화 적정결과와 그 화합물의 상태 살펴본 결과 금속이온이 0.2ml씩 더 참가되면서 색의 변화를 뚜렷하게 나타내었다. 반면 그 밖에 이온($Fe^{3+}$, $Ag^{2+}$, $Pd^{2+}$, $Zn^2$, $Fe^{2+}$, $Cu^{2+}$, $Pb^{2+}$)은 UV 흡광도 변화가 적거나 변화 자체가 없었다. 하지만 과량의 $Fe^{3+}$, $Ag^{2+}$, $Pd^{2+}$는 색상 변화를 나타내었으며,이와 같은 흡광도 변화는 금속에 따라 약간의 차이가 있지만, 420nm를 등흡수점으로 하여, 580nm의 파장 영역에 있는 흡수 밴드의 세기는 감소하는 반면 400nm 파장 영역에 있는 흡수 밴드의 세기가 증가하였다. 1,3-bisdicyanovinylindane 화합물은 다양한 생물계 및 환경계에서 요구되는 micro mol에서 milli mol 농도 영역의 $Hg^{2+}$ 이온의 선택적이고 민감한 검출과 정량적인 분석에 유용하게 사용될 수 있을 것이다.

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Simple Measurement of Livestock Foul Smells by Gas Detector Tube (검지관을 이용한 축산 악취 간이 측정방법 연구)

  • Jung, Jae-Chil;Ahn, Min-Ho;Jeong, Jong-Won;Choi, Hee-Seon;Kim, Tai-Jin
    • KSBB Journal
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    • v.24 no.5
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    • pp.420-424
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    • 2009
  • The present study was aimed for the fetor in the livestock facilities. Simple detection systems for ammonia and hydrogen sulfide were examined to see the mechanism making the discoloration length of malodorous substance concentration. Detector tube for ammonia showed a good linearity of 99.6% while hydrogen sulfide detector tube gave 99.7% linearity with reproductivity of 95%.

Analysis of Chemical Constituents of Saccharides and Triterpenoids in the Korean Native Mistletoes - II. Screening the Extractives of Korean Camellia Mistletoe (Pseudixus japonicus) for Cytotoxicity - (한국산(韓國産) 겨우살이류(類)의 당류(糖類)와 triterpenoids의 화학적(化學的) 조성(組成) 분석(分析) - II. 동백나무겨우살이 추출물의 항암활성 성분 검색 -)

  • Kim, Pyoung-Su;Ahn, Won-Yung
    • Journal of the Korean Wood Science and Technology
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    • v.24 no.1
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    • pp.87-94
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    • 1996
  • 한국산 동백나무겨우살이(Pseudixus japonicus) 추출물의 암치료를 위한 생약으로서 활성 유효성을 검증하기 위하여 메탄올, 석유에테르, 클로로포름, 초산 에틸 용매로 순차적으로 추출하여 겨우살이 추출물의 다섯 가지 분획을 얻어, 이에 대하여 in vitro로 1차와 2차 검색 시스템을 사용해 항암활성 성분을 체계적으로 검색하였다. 다섯 가지 분획 중 클로로포름 가용성 분획이 1차 검색 세포인 $P388D_1$에 대해 가장 높은 항암활성을 나타내어 MSB1, NIH/3T3, SNU-1, SNU-C2A 등 2차 검색 시스템에 대해 클로로포름 가용성 분획의 항암활성을 다양한 농도하에서 비교 검색하였다. 혈액암 세포중 특히 $P388D_1$의 생장이 클로로포름 추출물에 의해 강하게 저해되었으며, 형질전환된 생쥐의 태아 섬유아세포와 사람의 대장암, 위암세포들도 어느 정도의 생육저해를 나타내었다. 이 클로로포름 가용성 분획의 주성분은 원소분석, 발색시약과의 반응, IR, GC-MS, $^{13}C$-NMR의 스펙트럼의 결과로 세 종류의 알칼로이드 화합물로 확인되었고, 부성분으로는 지방산 메틸 에스테르와 프탈라이드 화합물이 MS 스펙트럼을 통해 동정되었다.

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Spot Test for Amins Acids with Alloxan (Alloxan 에 의한 Amino Acids 의 Spot Test)

  • Kim, Tae-Bong;Hahn, Bo-Sup
    • Journal of the Korean Chemical Society
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    • v.8 no.2
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    • pp.85-87
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    • 1964
  • In order to stabilze alloxan as a reagent for detection of amino acids by spot test, sugars and other reductants were added to the aqueous alloxan solution. It was found that lactose was the best for the purpose. The alloxan reagent containing lactose did not give color change on blank test and was very stable that there was no color change even it was allowed to stand in room temperature for several months. The color reaction with amino acids and some amines was not affected by lactose. This spot test for amino acids is in sensitivity as comparable to that of the previously reported methods and gave color reaction with proline and hydroxyproline to 1${\gamma}$ and 5${\gamma}$ respectively.

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An Enzyme-linked Immunosorbent Assay Strip Sensor for the Detection of Legionella Pneumophila (Legionella Pneumophila 검출을 위한 효소면역측정 스트립 센서)

  • Kim, Young-Kee;Park, Sojung
    • Applied Chemistry for Engineering
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    • v.25 no.5
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    • pp.544-547
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    • 2014
  • In this study, an enzyme-linked immunosorbent assay (ELISA) and immuno-chromatographic technique were combined to fabricate immuno-strip sensors for the detection of Legionella pneumophila. The immuno-strip sensor was manufactured with four different membranes. A nitrocellulose membrane was used to immobilize capture antibody and generate signals due to the high affinity to antibodies, and glass fiber membranes were used as a conjugate release pad and a sample application pad. A cellulose membrane was used as an absorption pad to induce sample flow by the capillarity. Colorimetric signals produced by sandwich immuno-reaction and enzyme reaction could be analyzed qualitatively and quantitatively within 30 min. Under the given experimental conditions, sensor signals with L. pneumophila samples were observed qualitatively by naked eyes and measured quantitatively in a range of $1.3{\times}10^3-1.3{\times}10^6CFU/mL$ with a digital camera and home-made image analysis software.